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"ABSTRAKAsam lipoat adalah senyawa yang terkandung dalam intra dan ekstraselular yang bertindak sebagai koenzim piruvat dehidrogenase, penangkal racun, pengkat logam berat dan antioksidan. Pengukuran asam lipoat diperlukan untuk mengetahui jumlah asam lipoat yang menjalankan fungsinya baik sebagai koenzim atau antioksidan. Tetapi pengukuran ini membutuhkan alat khusus seperti HPLC dan proses yang tidak mudah. Alat yang umum dan mudah seperti spektrofotometer diharapkan bisa menjadi alat penentuan asam lipoat. Pengukuran asam lipoat menggunakan spektrofotometri dengan metanol UV dan PdCl2 terlihat telah diuji dan diperoleh hasil yang valid dan dapat diandalkan dalam sediaan obat atau farmasi tetapi belum diuji untuk plasma. Pengukuran asam lipoat dalam plasma dan leukosit menggunakan PdCl2 menghasilkan pengukuran yang valid, dengan akurasi, presisi tinggi dan tidak berbeda dengan pengukuran asam lipoat menggunakan HPLC, p= 0,99. Sedangkan UV methanol berbeda dibanding HPLC p= 0,0001 atau tidak valid.

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ABSTRACTLipoic acid is a compound contained in intra and extracellular that act as a coenzyme of Pyruvate Dehydrogenase, antidote, chelating agent and antioxidant. Measurement of lipoic acid is needed to determine the amount of lipoic acid that performs its functions either as a coenzyme or an antioxidant. But this measurement requires a special tool such as High Performance Liquid Chromatography HPLC and a process that is not easy. A common and easy tool such as a spectrophotometer is expected to be a tool of lipoic acid determination. Measurement of lipoic acid using spectrophotometry with UV methanol and visible PdCl2 has been tested and obtained valid and reliable results in drug or pharmaceutical preparations but not yet tested for plasma. Determination of lipoic acid in plasma and leukocytes using PdCl2 produced valid result, with high accuracy, precision and was not different from lipoic acid measurement using HPLC, p 0.99. While UV methanol was different compare to HPLC p 0.0001 or was not valid."

"Keadaan hipoksia menyebabkan peningkatan Hypoxia Inducible Factor (HIP) sebagai respon terhadap menurunnya kadar oksigen. Selain menyebabkan peningkatan HIP, hipolsia juga menyebabkan peningkatan pembentukan dan penglepasan Reactive Oxygen Species (ROS) dari dalam mitokondria. yang kemudian akan meregulasi respons terhadap 02 yang rendah. Akibat peningkalan pembentukan ROS, kcmungkinan dapat teijadi stres oksidatif Penelitian ini bertujuan untuk mempelajari dan mengamati pengaruh hipoksia sistemik terhadap ekspresi gen HIF I-a dan stres oksidatif pada jaringan hati tikus yang diindiksi hipoksia sistemik selama I, 3, 7 dan 14 hari yang dibandingkan dengan kelompok normoksia sebagai kontrol.Induksi hipoksia sistemik dilakukan dengan memaparkan tikus jantan Sprague-Dawley terhadap lingkungan dengan oksigen l0% dan nitrogen 90% dalam sungkup hipoksiai Kada: protein, glutatzion (GSH) dan malondialdehid (MDA) diperiksa dari homogenat hati likus. Kadar protein dihitung dengan mengukur serapan pada 1 280 nm dan dibandingkan dengan serapan larutan standar Bovine Serum Albumin. Kadar malondialdehid (MDA) ditetapkan dengan metode Wiils dan kadar glutation (GSH) diukur dengan rnetode Ellman. Analisis ekspresi gen HIF 1-a dilakukan dengan metode Wesrern Blot dengan menggunakan anti HH? l-oi sebagai antibodi primer, anti IgG mouse sebagai antibodi sekunder dan pewamaan menggunakan aminoerhyl carbazole.Hasil penelitian menunjukkan bahwa kadar MDA hati meningkat mulai hari ke-l hipoksia dan bertahan sampai I4 hari, walaupun tidak bemiakna secara statistik Kadar GSH hati menunjukkan penutunan yang bermakna seiring dengan lamanya hipoksizi Hasil Weslerrz Blot menunjukkan adanya HIP I-a pada normoksia, hipoksia 1 hari dan 3 hari. Dapat disimpulkan bahwa terjadi stres oksidatif di jaringan hati seiring dengan lamanya hipoksia.

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Hypoxia condition increases the level of hypoxia-inducible factor (HIF) as response to oxygen deprivation. Hypoxia also increases production and releases of reactive oxygen species (ROS) from mitochondria Excessive production of ROS can lead to oxidative stress, due to its reactivity with macromolecules within cell, ie lipid. The objective of this study is to observe the effects of induction of systemic hypoxia on expression of HIP 1-c. gene and its relation oxidative stress in rat liver tissue.

The experiment was conducted on 25 male Sprague-Dawiey rats, which were divided into 5 groups : normoxic, hypoxia for l day, 3 days, 7 days and 14 days. Induction of systemic hypoxia was carried by exposing the rats in a hypoxic chamber with environment 10% 02 and 90% N2. To asses the oxidative stress condition, malondialdehyde (MDA) and glutation (GSH') concentration in liver was measured using Wills? and Ellman?s method, respectively. Expression of HIP 1-ot gene was analyzed using Westem Blot.

The result showed that MDA concentration is higher in all hypoxic group with no statistically significance difference. The GSH level decreased significantly until day 14. It seemed that oxidative stress occurred at day 14. HIF 1-a was expressed in normoxic condition, hypoxia day l and day 3. It was concluded that oxidative stress was more likely to occur at day 14 of hypoxia."

"ABSTRAKLatar Belakang: Komplikasi diabetes pada jantung dapat terjadi oleh karena produksi reactive oxygen spesies (ROS) berlebih. Beberapa studi menunjukkan stres oksidatif berperan dalam patogenesis komplikasi diabetes seperti kardiomiopati. Kurkumin telah terbukti memiliki khasiat sebagai antioksidan dan kardioprotektif. Tetapi kurkumin memiliki bioavailabilitas yang rendah didalam tubuh. Oleh karena itu kurkumin dibuat dalam bentuk nanokurkumin. Penelitian ini bertujuan untuk mengetahui pengaruh nanokurkumin terhadap stres oksidatif pada jantung tikus yang di induksi diabetes.Metode: Tikus Sprague-Dawley jantan di induksi diabetes dengan nikotinamide (NA) 100 mg/kgBB dan streptozotocin (STZ) 55 mg/kgBB secara intraperitoneal dan dosis tunggal. Terdapat 4 kelompok tikus antara lain, kelompok normal (tikus yang tidak di induksi), kelompok kontrol diabetes (CMC 0,5%), kelompok tikus diabetes yang diberi kurkumin oral 100mg/kg/hari dan kelompok tikus diabetes yang diberi nanokurkumin oral 100mg/kg/hari. Pengamatan dilakukan selama 30 hari. Kadar glukosa darah, aktivitas enzim creatine kinase myocardial band (CKMB), kadar malondialdehid (MDA), aktivitas enzim superoksida dismutase (SOD), glutation peroksidase (GPx) dan histopatologi otot jantung dianalisis dengan statistik menggunakan uji ANOVA, perbedaan dianggap bermakna secara statistik bila p<0.05.Hasil: Pemberian nanokurkumin dan kurkumin tidak mempengaruhi kadar glukosa darah dan cenderung menurunkan aktivitas CKMB pada serum. Nanokurkumin menurunkan kadar MDA jantung. Selain itu, nanokurkumin dan kurkumin dapat meningkatkan aktivitas enzim GPx tetapi tidak mempengaruhi aktivitas enzim SOD. Kurkumin memperbaiki kerusakan otot jantung dan lebih baik dibanding nanokurkumin.Kesimpulan: Hasil penelitian ini menunjukkan bahwa induksi diabetes dan pengamatan selama 30 hari belum memicu kondisi stres oksidatif yang nyata. Nanokurkumin tidak mampu memperbaiki kerusakan otot jantung tetapi mempunyai efek menekan kadar MDA dan meningkatkan aktivitas GPx.

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ABSTRACT

Background: Complications of diabetic in the heart may occur due to the excess production of reactive oxygen spesies (ROS). Previous studies showed that oxidative stress played a role in the pathogenesis of diabetic complications such as cardiomyopathy. Curcumin has potential and efficacy as an antioxidant and cardioprotective agent. However, curcumin has low bioavailability in the body. In the present study we investigate the effects of curcumin in the form of nanocurcumin against oxidative stress in the heart from streptozotocinnicotinamide- induced diabetic rats.

Methods: Sprague-Dawley rats were induced diabetes with nicotinamide 100mg/kg and streptozotocin55 mg/kg intraperitoneally. Rats were divided into nondiabetic group, diabetic control group (CMC 0,5 %) and two treated groups which were orally given curcumin at a dose of 100 mg/kg/day and nanocurcumin at a dose of 100 mg/kg/day, respectively. After 30 days of observation, the blood glucose levels, activity of the enzyme creatine kinase myocardial band (CKMB), levels of malondialdehyde (MDA), activity of the enzyme superoxide dismutase (SOD), glutathione peroxidase (GPx) and histopathology of the heart muscle were analyzed and the data were assessed using ANOVA test with the level of significancy of p <0.05.

Results: Nanocurcumin and curcumin did not decrease blood glucose levels and tended to reduce the activity of CKMB in serum. Nanocurcumin reduced cardiac MDA. Nanocurcumin and curcumin enhanced the activity of GPx enzyme, but did not influence the activity of SOD enzyme. Curcumin appeared to be able to repair injured heart muscle and was better than nanocurcumin.

Conclusion: The results of studyindicate that induction of diabetes by streptozotocin-nicotinamide did not result in severe oxidative stress in the rats. Nanocurcumin is not able to repair injured heart muscle but could suppress MDA levels and increase the activity of GPx."

"Latar Belakang. Penelitian ini bertujuan untuk membakukan teknik pengukuran asam folat serum menggunakan protein ikat folat PIF yang diisolasi dan dimurnikan secara utuh dari susu sapi segar dengan teknik enzyme labeled protein ligand binding assay ELPLBA . Metode. Metode yang digunakan adalah metode eksperimen yang menguji validitas dan perbandingan teknik ELPLBA untuk pembakuan teknik pengukuran folat serum.Hasil. Hasil isolasi dan pemurnian PIF menghasilkan kadar 3 tiga mg/mL. Identifikasi SDS-PAGE dan Western blot menunjukkan 3 tiga pita protein yang diperiksa adalah protein ikat folat. Validitas teknik uji keterulangan menunjukkan nilai yang dapat diterima CV.

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Background. This study was aimed to standarized technique of folic acid level serum measurment using folate binding protein FBP that isolated and purified from fresh bovine milk with enzyme labeled protein ligand binding assay ELPLBA technique. Method in this study, we performed an experiment research validated dan compared measurement techniques of ELPLBA with competitive ELISA for the standardization of the serum folic acid measurement technique. Results. FBP concentration yielded from isolated and purification was resulted 3 mg mL. SDS PAGE and western blot result showed 3 three protein bands that was confirmed to be FBP. Validity test repeatability indicate an acceptable CV 10 , whereas reproducible test showed poor results over a 5 day period. The results of the accuracy test of the enzyme labeled protein ligand binding assay technique showed good accuration. Linearity test of two samples showed quite linear results. Comparison of folic acid level measurement in serum between ELPLBA and ELISA technique showed there is no difference between both technique based on independent test T test at 95 confidence level. Conclusion. the enzyme labeled protein ligand binding assay technique on serum folic acid measurements were quite valid dan equivalent to the results obtained by competitive ELISA techniques."

"Tiamin (Vitamin B1) adalah vitamin B yang pertama kali diidentifikasi. Tiamin berperan sebagai koenzim untuk beberapa enzim yang terlibat dalam metabolisme energi.Uji laboratorium terhadap kekurangan tiamin dapat dilakukan dengan mengukur kadar tiamin dalam darah. Penelitian ini bertujuan untuk mengetahui kadar tiamin serum pada alkoholic dan penderita DM dengan teknik ELISA, HPLC, dan menggunakan protein ikat tiamin kacang hijau (PITKH) dengan teknik enzyme-labeled protein ligand assay (ELPLA). Untuk melihat faktor-faktor yang mempengaruhi ikatan antara PITKH dengan tiamin digunakan teknik dialisis kesetimbangan. Validitas teknik ELPLSBA dilakukan dengan uji presisi dan akurasi. Teknik ELISA dan HPLC digunakan sebagai pembanding pada pengukuran tiamin serum. Konsentrasi PITKH pasca kromatografi afinitas hasil pengenceran liofilisat stabil selama 30 hari pada suhu -20°C dan 3 hari pada suhu 4°C. Aktifitas pengikatan PITKH dengan tiamin optimum pada pH 7,5. Aktifitas pengikatan ini juga dipengaruhi oleh senyawa alkilasi, oksidator, dan reduktor, tetapi kurang dipengaruhi oleh ion kalsium dan logam-logam berat. Kemampuan PITKH dalam mengukur kadar tiamin serum dengan teknik ELPLA memiliki presisi dengan CV 4,1% dan akurasi dengan nilai R 96-98%. Pengukuran dengan ELISA memberikan hasil yang lebih rendah dari teknik ELPLA, sedangkan uji banding dengan HPLC diperoleh p = 0,102 (p > 0,05) ; artinya tidak terdapat perbedaan yang signifikan antara metode ELPLA dan HPLC. Pengukuran tiamin serum dengan teknik ELISA, HPLC dan ELPLA pada alkoholic dan penderita DM, lebih rendah dari serum normal.

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Thiamine (vitamin B1) was the first B vitamin to have been identified. It serves as a coenzyme for several enzymes involved in energy metabolism. The laboratory test against thiamine deficiency can be done by measuring thiamine levels in the blood. The aim of this study was to determine the serum thiamine levels in alcoholics and DM by ELISA, HPLC, and using mung bean thiamine binding protein (MBTBP) with the development of enzyme-labeled protein ligand assay (ELPLA) method. The equilibrium dialysis technique was used to see the factors affecting the bond between TBP and thiamine. The ELPLA method validity was performed with precision and accuracy tests. ELISA and HPLC methods were used as comparators for measurements of serum thiamine. The MBTBP concentration of post-chromatographic affinity resulted from dilution of lyophilisate was stable for 30 days at -20°C and 3 days at 4°C. The optimal pH for binding MBTBP to thiamine was 7,5. This binding activity was also affected by alkylation, oxidizing, and reducing agents, but it was less affected by calcium ions and heavy metals. MBTBP ability to measure serum thiamine levels with the ELPLA technique has precision with CV 4,2% and accuracy with R 96-98%. Measurements by ELISA has lower result than ELPLA. The comparison test with HPLC method obtained p = 0,102 (p > 0,05); meaning no significant difference between ELPSLBA and HPLC methods. Serum thiamine level by ELISA, HPLC and ELPLSBA techniques in alcoholic and DM patients were lower than normal serum."

"Latar belakang. Latihan fisik yang dijalankan secara teratur dengan intensitas dan durasi tertentu akan merangsang remodeling jantung sebagai usaha untuk mempertahankan fungsi ventrikel terhadap peningkatan beban biomekanik pada jantung. Diperkirakan bahwa latihan fisik jangka panjang menimbulkan remodeling jantung menyerupai hipertrofi kardiomiopati, berupa hipertrofi miosit dengan kekacauan tatanan miosit, fibrosis, apoptosis dan ko-lokalisasi gap junction. Tujuan penelitian. Mengetahui dampak latihan fisik jangka panjang dan henti-latih pada remodeling kardiomiosit. Metode penelitian. Penelitian eksperimental in vivo pada tikus Wistar ini dilakukan di Departemen Biokimia dan Biologi Molekuler, Laboratorium Imunohistologi Departemen Patologi Anatomi dan Bagian Fisiologi, FKUI. Latihan fisik dengan intensitas dan jangka waktu latihan yang berbeda, serta henti latih setelah periode latihan diterapkan pada tikus Wistar jantan dewasa muda. Dilakukan analisis terhadap perubahan morfologi kardiomiosit, fibrosis, apoptosis (ekspresi Caspase-3, Bax dan Bcl-2), gap junction (ekspresi Connexin43) dan pola EKG. Perubahan morfologi kardiomiosit diamati menggunakan pulasan Hematoxylin Eosin, fibrosis diamati menggunakan pulasan Masson?s Trichrome, sedangkan ekspresi Caspase-3, Bax, Bcl-2 dan Connexin43 diamati melalui pulasan imunohistokimia. Rekaman EKG dilakukan dengan filter 100 Hz, pada kecepatan kertas 50 mm/detik dan kepekaan 1 mV = 20 mm. Hasil dan pembahasan. Hasil penelitian ini menunjukkan bahwa latihan aerobik dan anaerobik menimbulkan hipertrofi eksentrik dengan peningkatan fibrosis dan apoptosis serta ko-lokalisasi gap junction ke arah lateral membran. Perubahan kardiomiosit, peningkatan fibrosis dan apoptosis lebih nyata pada latihan anaerobik dibandingkan latihan aerobik. Pola EKG menunjang adanya pembesaran ventrikel akibat latihan aerobik dan anaerobik disertai gangguan repolarisasi yang nyata terutama pada latihan anaerobik. Henti latih tidak mengembalikan morfologi miosit, apoptosis dan lokalisasi gap junction ke keadaan semula. Pola EKG setelah periode henti latih pada latihan aerobik tetap menunjukkan adanya hipertrofi ventrikel tanpa gangguan penghataran impuls yang berarti, namun pada latihan anaerobik tetap didapatkan gangguan repolarisasi berupa pemanjangan interval QTc yang bermakna. Kesimpulan. Remodeling kardiomiosit akibat latihan fisik jangka panjang tidak menyerupai struktur hipertrofi kardiomiopati, namun disertai peningkatan apoptosis, kolokalisasi Cx43 dan gangguan penghantaran impuls. Henti-latih tidak memulihkan remodeling jantung maupun gangguan penghantaran impuls listrik.

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Background. Regular physical exercise with certain intensity and duration stimulates remodeling of the heart as an effort to preserve ventricular function against an increased biomechanical load. It is postulated that long-term exercise induces cardiac remodeling that resemble cardiomyopathy hypertrophy marked by cardiomyocyte disarray, fibrosis, apoptosis and co-localization of gap junction. Research objective. To study the effect of long-term physical exercise and detraining on cardiomyocyte remodeling.

Methodology. This in vivo experimental study was conducted at the Departement of Biochemistry and Molecular Biology, Immunohistology Laboratory of Departement of Pathological Anatomy and Departement of Physiology FMUI. Physical exercise with different intensity and periods of training was performed in groups of young adult male Wistar rats, followed by a period of detraining. Analysis of cardiomyocyte morphological changes, fibrosis, apoptosis (expression of Caspase-3, Bax and Bcl-2), gap junctions (expression Connexin43) and ECG pattern was conducted. Changes in cardiomyocyte morphology was observed using Haematoxylin Eosin staining, fibrosis was observed using Masson's Trichrome staining, whereas the expression of Caspase-3, Bax, Bcl-2 and Connexin43 was observed through immunohistochemical staining. ECG recording was done with a filter of 100 Hz, the paper speed of 50 mm/sec and the sensitivity of 1 mV = 20 mm.

Results and discussion. The results showed that aerobic and anaerobic exercises cause the development of eccentric hypertrophy, with increased fibrosis and apoptosis as well as co-localization of gap junction to the lateral site of the membrane. Cardiomyocyte remodeling, fibrosis and apoptosis were more prominent in anaerobic compared to aerobic exercise group. The ECG pattern supports enlargement of the ventricles due to aerobic and anaerobic exercises with noticeable repolarization disturbances, especially in the anaerobic group. Detraining did not return myocyte morphology, apoptosis and localization of gap junctions to its basic state. The ECG pattern after a period of detraining following aerobic exercise supports the existence of ventricular hypertrophy without significant disturbances in impulse conduction, however repolarization disturbances in the form of significant prolongation of QTc interval persist in the group with anaerobic exercise.

Conclusion. Cardiomyocyte remodeling due to long-term physical exercise did not resemble cardiomyopathy hypertrophy structure, although increased apoptosis, colocalization of Cx43 and distrbances in impuls conduction were observed. Detraining did not restore cardiac remodeling and disturbances in electrical impulse conduction."

"Lingkungan mikro tumor berperan penting dalam meregulasi sifat kepuncaan, proliferasi, ketahanan terhadap apoptosis, dan metabolisme sel punca kanker. Penelitian ini bertujuan untuk menganalisis efek modulasi lingkungan ekstraseluler melalui kondisi hipoksia dan alkalinisasi pada metabolisme glukosa dan ketahanan hidup sel punca kanker CSC payudara manusia CD24-/CD44 . Pada penelitian in vitro eksperimental ini, CSC payudara manusia dikultur pada kondisi hipoksia dan kondisi alkali. Kultur sel diinkubasi selama 30 menit, 4, 6, 24, dan 48 jam pada suhu 37 C kemudian dilakukan analisis status metabolisme glukosa, regulasi pH, ketahanan hidup, dan penanda kepuncaan serta pluripotensi CSC payudara menggunakan berbagai teknik yaitu qRT-PCR, kolorimetri, fluorometri, dan aktivitas enzimatik. Kondisi hipoksia menyebabkan peningkatan ekspresi mRNA dan konsentrasi HIF1? sehingga mengaktivasi gen-gen yang berada di bawah regulasinya. Hipoksia juga menyebabkan penekanan proliferasi namun meningkatkan ketahanan terhadap apoptosis. Alkalinisasi menyebabkan peningkatan pH ekstraseluler pHe yang menstimulasi peningkatan aktivitas dan ekspresi mRNA gen regulator pH seluler. Status metabolisme menunjukkan peningkatan aktivitas glikolisis anaerobik disertai peningkatan ekspresi transporter GLUT1. Alkalinisasi menyebabkan penekanan proliferasi CSC payudara bahkan kematian sel. Sebagai kesimpulan, modulasi lingkungan ekstraseluler baik melalui hipoksia maupun alkalinisasi dapat meningkatkan aktivitas glikolisis yang selanjutnya mempengaruhi ketahanan hidup dan kepuncaan CSC payudara CD24-/CD44.

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This study was aimed to analyze the effect of extracellular pH and O2 level modulation on glucose metabolism and survival of the human CD24 CD44 breast cancer stem cells BCSCs . The primary BCSCs CD24 CD44 cells were cultured under hypoxia 1 O2 or under supplementation of sodium bicarbonate 100 mM for various periods. After each incubation periods, the pH regulation, glucose metabolism, survival, stemness and pluripotency markers were analyzed using various techniques including qRT PCR, colorimetry, fluorometry, dan enzymatic reactions. This study demonstrated that hypoxia caused an increase of HIF1 mRNA expression and protein level, and shifted metabolic states to be more glycolytic. Hypoxia also promoted the suppression of cell proliferation and induced the apoptosis evasion. Alkalinization caused a high pHe then stimulated an increase of mRNA and activity of cellular pH regulator that lead to upregulation of anaerobic glycolysis. Alkalinization inhibited BCSCs proliferation and promoted apoptosis. To conclude, modulation of the extracellular environment of human BCSCs through hypoxic condition and alkalinization could shift the metabolic state toward the anaerobic glycolysis which in turn affected the proliferation and survival."

"ABSTRAKLatar Belakang: Hipertrofi jantung dapat timbul akibat stres patologis misal hipoksia yang merupakan respon jantung sebagai mekanisme homeostatis yang diperlukan untuk menormalkan stres dinding ventrikel kiri dan mempertahankan curah jantung. Hipoksia sistemik kronik merupakan stres lingkungan yang berat. Respon spesifik jantung terhadap stres jantung terlihat pada peningkatan kadar peptida di dalam plasma, yang membantu jantung dalam menghadapi beban yang meningkat. Menurut sejumlah peneliti, kadar Apelin berhubungan erat dengan disfungsi ventrikel. Apelin merupakan preproprotein dengan 77 asam amino yang disekresikan dari keluarga adipokine, berperan dalam mempertahankan performa jantung pada beban tekanan kronik. Pada tingkat molekular, respons adaptasi diperantarai oleh perubahan ekspresi gen. Tujuan penelitian: Menganalisis pola ekspresi gen Apelin dan gen BNP pada hipertrofi ventrikel akibat induksi hipoksia sistemik kronik dengan mengukur konsentrasi Apelin-13 dan konsentrasi BNP-45. Penelitian bersifat eksperimental menggunakan 28 ekor tikus Sprague-Dawley jantan, umur 8-12 minggu yang dibagi dalam 7 kelompok n=4 ekor/kelompok , terdiri dari kelompok kontrol normoksia, O2 atmosfir dan kelompok perlakuan hipoksia dalam sungkuphipoksia, 8 O2, masing-masing selama 6 jam, 1, 3, 5, 7 dan 14 hari . Parameter stres oksidatif akibat hipoksia jantung, dilakukan dengan pengukuran kadar malondialdehid MDA dan histopatologi dengan pewarnaan HE. Selain itu juga dilakukan pengukuran protein Apelin-13 dan BNP-45 menggunakan metoda ELISA dan pengukuran ekspresi relatif mRNA Apelin dan BNP-45 jantung, menggunakan real time RT-PCR kuantitatif dengan rumus Livak. Hasil penelitian: ekspresi relatif Apelin-13 di jantung menurun pada awal hipoksia dan kemudian meningkat mulai hari ke-3 sampai hari ke-14. Peningkatan kadar MDA yang signifikan terjadi sejak hipoksia 7 hari. Korelasi MDA terhadap peningkatan ekspresi relatif Apelin adalah kuat r=0.750 dan signifikan p=0.000 . Korelasi BNP-45 terhadap Apelin-13 adalah sangat kuat r=0.943 dan signifikan p=0.000 . Dapat disimpulkan bahwa adanya peningkatan MDA, peningkatan ekspresi relatif dan protein Apelin-13 dan peningkatan ekspresi relatif dan protein BNP-45 pada jaringan jantung mempunyai korelasi yang signifikan dan kuat, sesuai dengan peningkatan lamanya perlakuan hipoksia.

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ABSTRACTBackground: Cardiac hypertrophy can result from pathological stress eg hypoxia as a response to ventricular wall stress and to maintain cardiac output. Chronic systemic hypoxia is a severe environmental stress. During cardiac stress certain peptides are release by the heart into the plasma, which help the heart to compensate the increased myocardial load. According to several authors, apelin levels are increased during cardiac dysfunction. Apelin is a preproprotein with 77 amino acids from adipokine, which contributes to maintaining cardiac performance at chronic stress loads. At the molecular level, the adaptation response is mediated by changes in gene expression. Objective: To analyze the expression pattern of Apelin-13 and BNP-45 on ventricular hypertrophy due to induction of chronic systemic hypoxia by measuring Apelin-13 and BNP-45 concentrations. The experimental study used 28 male Sprague-Dawley rats, 8-12 weeks old divided into 7 groups 4 per group , consisting of control group normoxia, atmospheric O2 and 4 hypoxia treatment groups, which underwent systemic hypoxia in hypoxic chamber containing 8 oxygen, respectively for 6 hours, 1, 3, 5, 7 and 14 days . The presence of oxidative stress due to cardiac hypoxia was determined by malondialdehyde MDA and cardiac structural alteration was examined by HE staining. Apelin-13 and BNP-45 proteins were determined using the ELISA method and the relative expression of cardiac Apelin and BNP-45 mRNA were determined using quantitative RT-PCR real time with Livak formula. Results: Relative expression of Apelin-13 in the heart decreased early in hypoxia and then increased from day 3 to day 14. Significant increases in MDA levels occurred after 7 days hypoxia. There was a strong and significant correlation between MDA levels and Apelin relative expression r = 0.750, p = 0.001 . Similar results were obtained for of BNP-45 and Apelin-13 r = 0.943, p = 0.001 . From the results, it can be concluded that during chronic systemic hypoxia there was an increase in oxidative stress, relative expression and Apelin-13 proteins and relative expression and BNP-45 protein of the rat cardiac tissue."